Värmechockprotein 90 kDa (HSP90) bildar ett anmärkningsvärt komplicerat nätverk med en mängd olika cochaperones. Komplexet av FK506-bindande protein 51 kDa (FKBP51) och HSP90 förmedlar proteinveckning och funktion, främjar tau aggregation vid Alzheimers sjukdom och påverkar stressrelaterade störningar, fetma, typ två-diabetes, etc. I samarbete med den molekylära chaperonen HSP90, FKBP51 har nyligen föreslagits som ett lovande terapeutiskt mål för Alzheimers sjukdom (AD). Således skapades knock-in-musen med punktmutationer i tetratricopeptide repeat (TPR) domänen av FKBP51, vilket gör den oförmögen att interagera med HSP90, för att undersöka de potentiella terapeutiska målen för behandling av dessa sjukdomar. Glukokortikoidreceptorn (GR) fungerade traditionellt som utgångspunkten för de initiala studierna av FKBP51-funktion och mekanism som kan stimuleras av den syntetiska glukokortikoiden dexametason (Dexa). Det primära målet med projektet är att förstå den biologiska betydelsen av FKBP51-HSP90 interaktioner. Det är oklart hur FKBP51-mutation påverkar protein-protein-interaktionen och glukokortikoidsignalering. Här analyserades embryonala fibroblaster (MEF) isolerade från vildtyp och FKBP51 mutant mus med avseende på proteinlokalisering, proteinuttryck och genuttryck. Även om ingen säker skillnad mellan vildtyp och mutantmöss sågs i Dexa-medierad glukokortikoidsignalering, förekommer de posttranslationella modifieringarna (PTM) vid exponering för Dexa-behandling av FKBP51 i vildtypmöss i en signifikant högre utsträckning än i Fkbp51mute-möss.Fosforyleringsmodifieringen av FKBP51 antogs initialt och bekräftades av fosforyleringsanrikningsstrategier. Bekräftelse har dock ännu inte erhållits. / Heat shock protein 90 kDa (HSP90) forms a remarkably complicated network with a variety of cochaperones. The complex of FK506-binding protein 51 kDa (FKBP51) and HSP90 mediates protein folding and function, promoting tau aggregation in Alzheimer's disease and influencing stress-related disorders, obesity, type two diabetes, etc. In collaboration with the molecular chaperone HSP90, FKBP51 has recently been proposed as a promising therapeutic target for Alzheimer's disease (AD). Thus, the knock-in mouse harboring point mutations in the tetratricopeptide repeat (TPR) domain of FKBP51 rendering it unable to interact with HSP90 were created to investigate the potential therapeutic targets for the treatment of these diseases. Glucocorticoid receptor (GR) traditionally served as the starting point for the initial studies of FKBP51 function and mechanism which can be stimulated by the synthetic glucocorticoid, dexamethasone (Dexa). The primary goal of the project is to comprehend the biological significance of FKBP51-HSP90 interactions. It is unclear how FKBP51 mutation affects the protein-protein interaction and glucocorticoid signaling. Here, embryonic fibroblasts (MEFs) isolated from wildtype and FKBP51 mutant mouse were analyzed with respect to protein localization, protein expression, and gene expression. Although no certain difference between wildtype and mutant mice was seen in Dexa-mediated glucocorticoid signaling, the post-translational modifications (PTMs) in exposure to Dexa treatment of FKBP51 occur in wildtype mice to a significantly higher extent than in Fkbp51mute mice. The phosphorylation modification of FKBP51 was initially hypothesized and confirmed by phosphorylation enrichment strategies. However, confirmation has not yet been obtained.

<p>  </p> <p>Cilia are cell appendages that sense our environment and are critical in cell-to-cell communication. Dysfunction of cilia can result in several disease states including obesity. While cilia in the brain are known to be important for feeding behavior, it is unclear how they regulate energy homeostasis. Classically, cilia coordinate signaling through surface receptors called G-protein coupled receptors (GPCRs). For example, cilia mediated GPCR signaling is critical for both our senses of vision and smell. How cilia regulate the signaling of GPCRs in other areas of the body including the brain is only now emerging. To answer cell biology questions around cilia mediated GPCR signaling in neurons, we developed a system for primary neuronal cultures. We discovered that the cilia mediated hedgehog pathway influences the ability of neurons to respond to GPCR ligands. For the first time, this result highlights the role of the hedgehog pathway in neurons. We continue to explore how cilia integrate the hedgehog pathway and GPCR signaling in the central nervous system, and the potential connections to energy homeostasis. We discovered that hedgehog pathway activity in feeding centers of the brain changes based upon feeding conditions like fasting. We also learned that activating the hedgehog pathway in these brain regions is sufficient to cause obesity in mice. These novel results highlight an unrecognized role for the hedgehog pathway in the regulation of feeding behavior. Overall, this work provides a better understanding of ciliopathy associated obesity and may reveal more common mechanisms of obesity in the general population. In addition, this work implicates the hedgehog pathway in regulating behaviors and new modes of cell-cell communication within the central nervous system.</p>